Atraumatically formed tissue compositions, devices and methods of preparation and treatment

ABSTRACT

A process and system provides for atraumatic preparation of morcelized Tissue Particles (MTPs), such as Full Thickness Skin Graft Particles (FTSGPs), cartilage particles and other organ tissue particles, in a liquid medium. The resultant tissue product may be a suspension of Tissue Particles in an aqueous solution and containing highly viable cells and may be rapidly prepared at bedside or in the operating room and conveniently delivered to a patient through a syringe or similar applicator. The MTPs may be used for surgical applications including wound healing, cosmetic surgery, and orthopedic cartilage repairs.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation-in-part of U.S. patent applicationSer. No. 16/584,755, filed Sep. 26, 2019, which claims priority to U.S.Provisional Patent Application No. 62/843,724 filed on May 6, 2019, andU.S. Provisional Patent Application No. 62/844,232, filed May 7, 2019,and is a continuation-in-part of PCT International Application No.PCT/US2020/031286, filed May 4, 2020, which is a continuation-in-part ofU.S. patent application Ser. No. 16/584,755, filed Sep. 26, 2019, whichclaims priority to U.S. Provisional Patent Application No. 62/843,724filed on May 6, 2019, and U.S. Provisional Patent Application No.62/844,232, filed May 7, 2019, the contents of all of which areincorporated herein by reference in their entirety.

FIELD OF INVENTION

The present invention relates to a process, method, device, and systemfor preparing atraumatically finely Morcelized Tissue Particles (MTPs)in a liquid medium. The term Tissue Particles (TP) is meant to includetissue harvested from Full Thickness Skin Grafts (FTSGs), Split SkinGrafts (SSGs), Cartilage Grafts (CG), or other organ graft tissues,which is then subjected to the processes described herein to producesmall, morcelized tissue particles which retain their viability duringthe process of morcelization. The resultant product of this process maybe a suspension of the MTPs, such as full thickness skin graft orcartilage graft particulates, containing highly viable interconnectedtissue cells and extracellular matrix material, in an aqueoussuspension, which may be prepared rapidly in a closed aseptic system atbedside, as an office procedure, or in the operating room and deliveredconveniently and uniformly to treat a body wound/injury/defect through asyringe or with other controllable application methods. The TP, such asFTSGs, CG suspensions may be used as appropriately indicated in plasticsurgery, orthopedic surgery, or other surgical applications, such aswound healing, cosmetic surgery, or joint surgery, among others. Theterm MTPs is meant to refer to the finely cut, highly viable tissue asprocessed in accordance with the present invention, including varioussizes and shapes, for the purposes described herein. In the case of skinthe MTPs are desirably FTSGs.

BACKGROUND OF THE INVENTION

There are a number of challenges to overcome in processing tissues forgraft applications. For the treatment of skin wounds, the mostefficacious transplants are full-thickness skin grafts. Full-thicknessskin grafts involve the epidermis and the entire dermis as well,allowing for most of the characteristics of the grafted skin to bepreserved in the process. However since for transplanting FTSGs, theentire dermal layer needs to be removed, the resulting skin graft donorsite will not support regeneration and will need to be sutured closed.Consequently, only small FTSGs can be used by current methods.Nevertheless, the result of FTSGs is a graft that maintains more of thenormal characteristics of the skin (notably texture, color andthickness), and is also less likely to contract as it heals. This makesFTSGs the more aesthetically pleasing choice for grafts.

Alternatively, in order to treat larger wound areas, physicians usesplit-thickness skin grafts (STSG) because the resulting donor sites areable to heal on their own over time with appropriate wound dressings.STSG involve only the epidermis and variable portions of the dermis,leaving behind enough of the dermis for the donor site to heal byre-epithelialization without the need to close the donor site wound withsutures. However, these donor sites are painful and slow to heal and theSTSG grafts harvested are often still not sufficiently large to coverlarge wound areas. STSGs can then be meshed, allowing for smallersections of tissue to be expanded to effectively cover larger areas. Thecombination of meshed appearance, varying pigmentation, and thinnessmakes STSGs less cosmetically appealing than FTSGs. Most STSG proceduresmust be performed in operating room setting.

A variety of techniques and systems to effectively treat large wounds,improve donor site healing and also enable the procedure to be performedas an outpatient office procedure have been developed for various typesfor surgical applications.

Some techniques have focused on harvesting only the epidermis, ratherthan including the split or full thickness dermis. Using only theepidermis, the top outermost layer of skin, as a grafting technique hasits applications but provides the most limited grafting effectivenessbecause it contains none of the structural components or cells of thedermis that are desirable for improved healing.

One such commercially known device for harvesting epidermal tissue isthe Cellutome Epidermal Harvesting System, marketed by KCI, an AcelityCompany. This system uses a vacuum to create series of epidermalmicro-domes on the donor site which are cut by hand using a blade andtransferred to the patient using an adhesive pad. This technique can beperformed outside of the operating room and provides only forty-twosmall epidermal skin patches which are only suitable for graftingsuperficial wounds in small areas. Because these epidermal grafts do notcontain the structural components or cells of the dermis, these graftswork best on superficial wounds and results in small, superficial donorsites that require healing, albeit faster than donor sites from STSGs.

A recent variation of split thickness skin grafting technique is theXpansion Micro Auto Grafting Kit, marketed by Acell, Inc. This deviceconsists of disposable instruments designed to be used for theharvesting, mechanical preparation, and application of split-thicknessskin autografts for the purpose of transplantation onto wounds. In acommercial setting, this technique involves manually harvesting a smallsplit thickness skin graft with a hand dermatome type device and thenfurther mincing the graft by hand using a series of parallel cuttingdisk blades, and then spreading the minced pieces over a larger woundarea by using a spatula. This process still results in a donor site inneed of healing and also presents challenges with need to dislodge theminced tissue from between the stacked parallel roller blades and withthe handling and transfer or the small pieces of graft.

A recent variation of full thickness skin grafting technique isdescribed in the US Publications 2016/0310159 and 2016/0310157 as aharvesting device that processes full thickness grafts. This deviceutilizes rows or arrays of adjacent hollow needle-point tubes, assistedwith ultrasonics to core and capture tubular micro-columns of fullthickness skin samples from a donor site. The tubular columns of tissuearrays are intended to be scattered over a wound site (graft site). Thistechnique requires a large number of harvested tissue micro-columns,however the device is limited by the finite number of columns achievablewith each use. Also, like the Cellutome or Xpansion devices, this devicecreates an ancillary donor site in need of healing and the resultingtissue form is challenging to spread uniformly over irregular woundsites and is unsuitable for grafting large areas.

Until now, the means to effectively use some grafts for transplantationin surgical applications such as full thickness skin grafts, splitthickness skin grafts or cartilage grafts has remained inadequatelyresolved. The un-solved problems of how-to best process tissues, such asskin grafts or cartilage grafts in an effective manner by known priorart methods and devices, include among others: 1) the inability toharvest a large full thickness skin graft without creating an ancillarywound that is too large thus limiting FTSG, the gold standards for skingrafts, to very small grafts that can be closed with sutures; 2)creation of donor sites that are painful and slow to heal with STSGs; 3)and the inability to achieve a graft tissue forms from skin grafts orcartilage grafts with high cell viability that are easy to processrapidly, manipulate in a closed aseptic system, and to be applieduniformly over wounds with irregular surfaces; and 4) the inability toprocess full thickness skin grafts in a time and cost effective mannerthat can be expanded to cover larger areas or processed to treat chronicor contaminated wounds.

SUMMARY OF THE INVENTION

The present disclosure addresses many of the aforementioned issues oflive tissue processing. The device and processing methods arespecifically designed to process tissue, harvested a-traumatically intoparticulates in an aqueous suspension with very high cell viability thatcan be easily dispensed on a wound. Using full thickness skin containingall the skin cell types and skin extracellular matrix, the graft can beharvested from an ancillary site with the ability to completely sutureclose the donor site during the procedure and then process the tissue tocover an area much larger than the original skin area with a graftcontaining high viability autologous skin cells. The closed systemdevice and the aqueous environment allows for convenience, ease oftransfer, and control of sterility, temperature, and pH, withoutdetrimental loss of viability of the tissue cells in skin or cartilagegrafts. The resulting graft form is a liquid or paste form and can bedispensed precisely and uniformly as desired. In the case or cartilage,grafts can be taken from areas of non-articulation, then processed andgrafted into cartilage defects.

The resulting abundance of readily morcelized tissue form, in theexample of full thickness skin, containing all tissue components of theskin, that is a mixture of viable dermal and subdermal cells andinterstitial tissue components, exceeds the capability of othercurrently available devices and practiced methods.

Thus, in one aspect of the invention there is provided Full ThicknessSkin Graft (FTSG) composition comprising a plurality of mechanicallyseparated Full Thickness Skin Graft Particulates (FTSGPs) present in aliquid medium. The composition includes FTSGPs which include fullthickness skin cells and extracellular matrix material. The majority ofthe plurality of skin cells within the FTSGPs are desirably viable afterprocessing, with at least about 50% of the skin cells within FTSGPsviable after processing.

The FTSGPs have an average size as measured across their largestdimension of about 200μ to 1500μ (0.020 mm to 1.50 mm), desirably about350μ to 1250μ (0.35 mm to 1.25 mm) and more desirably about 500μ toabout 1000μ, and even more desirably about 500 to about 750μ or 250μ to750μ, relative to particular surgical implant applications. The nominalaverage size of tissue morcels may be controllably varied with durationand/or speed of mechanical processing, as desired for particularsurgical implantation purposes. The overall process is capable of beingexpediently completed to optimally maximize cellular viability of thetissue graft material from the time of harvesting to the time ofautologous implantation. The process of morcelization may be effectivelycompleted, for example, within three to ten minutes, relative to thetype of tissue being processed and the nominal tissue particle sizesdesired for a particular surgical application.

The TP, or in the case of skin, (e.g., FTSGPs, STSGPs) or cartilage(CPs), may be formed by atraumatically slicing the tissue intoparticulates in a liquid medium using the devices as further describedherein. Desirably, the liquid medium may be a hydrophilic medium, butmay also be an oleophilic medium. The MTPs (e.g., FTSGPs) may besuspended in the liquid medium. The liquid medium itself may be in theform of solution, an emulsion, a suspension and combinations thereof.

The present invention would process tissue that would include all thecell types and extracellular matrix components of the processed tissue.For full thickness skin, this would include all epidermal and dermalcells as well as skin appendage cells and extracellular matrix.

In another aspect of the invention there is included a device forprocessing organ tissue which includes:

-   -   a) a container for accommodating fluid and for receiving said        tissue;    -   b) a pair of cutting devices supported in juxtaposition in a        container, at least one of the cutting devices being moveable        with respect the other cutting device to slice tissue between        thereof; and    -   c) an agitation device which causes repeated flow of fluid and        fluid suspended tissue through the juxtaposed cutting devices,        with the agitation device capable of moving in concert with at        least one moveable cutting devices and repeatedly moving fluid        and tissue through the juxtaposed cutting devices repeatedly.

At least one movable cutting device may be adjacent to the other cuttingdevice. The movable cutting device may be mounted to or integrated withthe agitation device for movement. One of the juxtaposed cutting devicesmay be fixed.

The agitation device causes circulating flow of fluid and tissue throughthe juxtaposed cutting devices. Desirably, the agitation device createsa vortex propulsion movement of fluid and tissue repeatedly through thespace between the juxtaposed cutting blades. The agitation device maytake a variety of forms, desirably the agitation device includes or isan impeller for causing circulating flow of fluid and tissue repeatedlythrough the juxtaposed cutting blades. Moreover, it is also desirablethat the agitation device and the movable blade be movable in arotational direction about an axis.

As will be seen from the figures and descriptions, the impellerdesirably has a curved surface along said axis for causing continuouscirculating movement of liquid and tissue. Moreover, at least one ofsaid cutting devices includes blades radially arrayed with angularspaces between, and desirably the other cutting device includes aplurality of said blades radially arrayed with curved angular spacesbetween. The angular spaces between the blade edges of the movablecutting device may be different from the angular spaces of the blades ofthe juxtaposed cutting device. The cutting devices may be in physicalcontact along a common shear plane.

As described, the container includes an opening for receiving of fluidand tissue and may also include an outlet for the discharge of fluid andprocessed tissue. The system may also include a dispensing device influid communication with the outlet to receive the discharged fluid andthe processed tissue. The dispenser may be selected from a variety ofdevices, including syringes, which are particularly useful because it isan accurate and easy way to dispense the fluid composition containingmorcelized Tissue Particles or specifically FTSGPs onto a wound area.

As mentioned above, the present disclosure further includes a method forforming processed tissue into particulate form which includes:

providing a container supporting a pair of cutting devices injuxtaposition;

placing fluid and organ tissue into said container;

moving at least one of said cutting devices to cut said organ tissue;and

providing an agitation device to continuously move fluid and tissuethrough the cutting devices to repeatedly slice or cut tissue intoprogressively smaller particulates.

The method may further include providing a dispensing device in fluidcommunication with an outlet; and discharging the fluid and tissue intoa dispensing device.

In another aspect of the disclosure there is included a plurality ofMTPs, e.g., full thickness skin graft particulates (FTSGPs), desirablyin an aqueous suspension made by the process which includes:

-   -   providing a container supporting a pair of cutting devices in        juxtaposition;    -   placing fluid and tissue into the container;    -   moving at least one cutting device to cut tissue or in one case        FTSG tissue;    -   and providing an agitation device to continuously move said        fluid and said organ tissue through the cutting devices to        repeatedly tissue into progressively smaller particulates.

As previously mentioned, in such a product containing a plurality ofTissue Particles (TP) or in one case full thickness skin graftparticulates (FTSGPs), desirably the particulates are processed anddispensed in an aqueous suspension, and the majority of the plurality ofFTSGPs are viable after processing, desirably at least 50% or more. Thishigh viability is due to a number of factors, including the use ofatraumatic slicing by the inventive devices, the use of a liquid,biologically friendly medium, such as a saline or other isotoniccompatible medium which can buffer Ph, and prevent desiccation. Aspreviously mentioned, the choice of liquid medium may be one ofhydrophilic character, oleophilic character or may have aspects of both,such as an emulsion. The process also does not generate excessive heatand temperature can be controlled such that cells are negativelyaffected. Although various sizes of morcelization are contemplated, insome embodiments, the TP or FTSGPs have an average size as measuredacross their largest dimension of about 150μ to about 1000μ.

Smaller particle sizes facilitate dispensing through devices such assyringes, which are both familiar to the practitioner and easilymanipulated for controlled deposition at the wound site. For example,particles which are nominally smaller than 400μ are useful for deliverythrough an 18 gauge needle or nominally less than 200μ to be deliveredthrough a 22 gauge needle.

For subdermal implantation through needle injection, the epidermis isremoved prior to injection. This can be done through several methods andis a routine surgical procedure. With the epidermis removed only thedermal elements are processed and only the dermal elements without theepidermis can be injected into the dermal or subdermal plane. Thisoverall process is otherwise identical to the processing of other tissueor full thickness skin.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1-4 are sectional views showing of the processing device formorcelizing tissue grafts full thickness skin grafts (FTSGs).

FIGS. 5-8 show in schematic detail the morcelizing mechanism of theprocessing device of FIGS. 1-4.

FIGS. 9 and 10 show in further detail the morcelizing mechanism of thepresent invention.

FIG. 11A is a partial top view of a portion of the morceling mechanismof FIGS. 9 and 10.

FIG. 11 B is a side view, partially in section, of the morcelingmechanism of FIGS. 9 and 10.

FIGS. 12A-12D show various configurations for the blade edges and therelationship of the blades to a shear plane of the morcelizing mechanismof the present invention.

FIGS. 13A-13C show an alternate arrangement of the cutting blades of themorcelizing mechanism of the present invention.

FIG. 14 is a schematic representation of a recirculating flow pathcreated using the morcelizing mechanism of the present invention.

FIG. 15A is a sectional showing of the processing device of FIGS. 1-4attached to a preferred embodiment of an applicator.

FIG. 15B is an exploded view of the applicator of FIG. 15A.

FIGS. 15C and D are side plan and section views, respectively, of theapplicator of FIG. 15B.

FIG. 15 E is a horizontal section view of the applicator of FIG. 15B.

FIGS. 16-18 shown in partial section are further embodiments of theprocessing device of the present invention attached to furtherembodiments of isolation and applicator devices.

FIG. 19 schematically shows a processing device in conjunction with afurther embodiment of a removable isolation device which is configuredfor use as an applicator device.

FIGS. 20 and 21 show axial and lateral sectional views of the processingdevice of the present invention including baffles to divert a singlelarge vortex into separate smaller vortexes to more expedientlymorcelize tissue.

FIG. 22 shows, in section, the flow through the processing device of thepresent invention which recirculates externally from the processingdevice container.

FIG. 23 is a schematic diagram showing a processing device of thepresent invention used in conjunction with a centrifugal device toisolate and compact particles from solution.

FIG. 24 is a schematic flow diagram of the system progression of thepresent invention.

FIG. 25 is a table showing samples of FTSGPs which were processedaccording to the invention and exhibited cell viability of 87.7-98.1%during and immediately after the processing using the inventive devices,methods and systems.

FIG. 26A shows a photograph of an inventive sample of morcelized FTSGPsin fluid suspension drawn from an inventive processor as describedherein without baffles after 4 minutes of processing.

FIG. 26B shows a photograph of a subsequent inventive sample ofmorcelized FTSGPs drawn after a total of 7 minutes of processing withoutbaffles.

FIG. 27 shows a photograph of an inventive sample of morcelized FTSGPsusing an inventive process and device without baffles after a total of 7minutes of processing.

FIG. 28A is a close-up view of a portion of the FTSG prior toprocessing, with a sectional view revealing the thin layer of epidermaltissue (typically including pigmented stratum corneum, stratum lucidum,statum granulosum, thickly cell populated stratum spinosum, and stratumbasale), over the thicker layer of generally white dermis (includingdermal papilla, stem cell rich hair follicles, sweat glands,capillaries, sensory nerve fibers, sebaceous glands and other dermalcomponents—all contained within an abundance of collagen fibers andconnective tissue).

FIG. 28B shows the resultant dense mixture of inventive morcelizedFTSGPs particles suspended in 35 ml of buffer solution, contained withinthe processor chamber.

FIG. 28C shows an enlarged view of the densely populated inventive FTSGPtissue particle solution presented in a petri dish.

FIG. 28D shows an enlarged view of the inventive processed MTPs,(FTSGPs) annotated to point out that the mixture contains differingamounts of epidermis (pigmented) versus dermis (generally whiter),varying proportionally looking at the sectional view of pre-processedtissue.

FIGS. 29A-E show samples processed using the same device and sameparameters as FIGS. 28A-D, on a different day and with abdominoplastyderived from a different patient.

FIGS. 30A-B shows harvested cartilage portions and morcelized cartilagerespectively in accordance with the present invention.

FIG. 31 shows schematically the device technology used in accordancewith the present invention including a processing device, an applicatordevice and reusable equipment.

FIG. 32 shows sequentially, the preparation, processing and applicationemployed in accordance with the present invention.

FIG. 33 shows the present invention with reference to full thicknessskin grafts and cartilage grafts.

FIG. 34 shows the process employed in in the present invention includingintroduction, morcelization and dispensing.

FIG. 35 shows variable tissue particle size produced in accordance withthe present invention including full thickness skin graft particlescontaining cells and extracellular connective tissue.

FIG. 36 shows dispensing options which may be employed in accordancewith the present invention.

FIG. 37 shows the examples of clinical indications for the presentinvention.

FIG. 38 shows the current developmental status of the present invention.

FIGS. 39-41 are schematic vertical cross sections of a furtherembodiment of the processing device of the present invention eachshowing successive stages of operation.

FIGS. 42 and 43 are schematic cross sections of a still furtherembodiment of the present invention each showing successive stages ofoperation.

FIGS. 44 and 45 are schematic vertical cross sections of a furtherembodiment of the processing device of the present invention eachshowing successive stages of operation.

FIG. 46 is a schematic vertical cross section of a still furtherprocessing device of the present invention.

FIG. 47 is an enlarged portion of the embodiment of FIG. 46 showingsuccessive operation.

FIG. 48 is a horizontal sectional showing of a filter chamber of thepresent invention.

FIGS. 49 and 50 shown in horizontal cross section and perspective,respectively the processing chamber of the present invention withbaffles shown therein.

FIGS. 51 and 51A are exploded perspective and side views respectively ofthe cutting assembly and impeller of the present invention.

FIG. 52 is a rear perspective view of the cutting assembly of FIG. 51.

FIGS. 53 and 54 are side plan and exploded perspective showingsrespectively of a further embodiment of the cutting assembly of thepresent invention.

FIG. 55 is a side plan showing of a still further embodiment of thecutting assembly of the present invention.

FIGS. 56-58 show output of FTSGs processed at various output volume.

FIG. 59 shows an excised elliptically shaped FTSG.

FIG. 60 shows processed FTSGPs in a petri dish processed from the FTSGof FIG. 59.

FIG. 61 shows another excised elliptically shaped FTSG.

FIG. 62 shows processed FTSGPs in a petri dish processed from the FTSGof FIG. 61.

FIG. 63 shows finely morselated cartilage in a petri dish.

FIG. 64 shows a planar slice of morselized cartilage using fluorescentconfocal microscopy.

DETAILED DESCRIPTION OF THE INVENTION

The present invention enables the processing of excised tissue biopsysamples (TBSs) into a multitude of morcelized tissue particles (MTPs),for autologous wound healing. MTPs achieve effective healing byfacilitating rapid cellular outgrowth from viable cells within amultitude of micrograft particles to radiate peripherally from and tobridge between the individual micrografts applied upon and covering anopen wound.

The present invention embodies a sterile single-use disposable devicewhich enables an effective and expedient means for a surgeon or medicalpractitioner to aseptically process an excised biopsy into an abundanceof autologous micrograft particles. The process can be performed in anoperating room or in a clinic at a patient's bedside. The procedure canbe performed within a single patient visit procedure.

TBSs are processed into MTPs for example, full thickness skin graftparticles (FTSGPs), split thickness skin graft particles (“STSGPs”) andcartilage particles (“CPs”), in a fluid suspension for delivery to theautologous treatment area. This can be viewed as different stages, forexample:

Stage 1 Harvesting

The first stage is to taking a tissue biopsy sample (TB S) from apatient for the purpose of processing the sample in accordance with theinvention, into a morcelized form which can be applied to the area to begrafted (treated). A non-limiting example of a TBS may be an abdominaltissue biopsy sample, which can be taken and processed according to theinvention. Other types of tissue biopsy samples may of course be taken,from different areas of the body and from different tissue types,including but not limited to cartilage graft biopsy samples, fullthickness skin graft biopsy samples and split thickness skin graftbiopsy samples. All such samples are included in the definition of TBS.These biopsy samples are relatively large compared to what they will beonce they are morcelized and processed according to the invention.

Stage 2 Set-Up/Preparation

The TBS are added to a solution, such as saline solution, in theprocessing chamber of the system of the present invention. Desirably thesolution is a sterile solution, e.g. sterile saline, or a solutionbuffered to a pH that is favorable to the tissue type and viability ofthe TBS.

Stage 3 Processing

The TBS are processed in the solution. The morcelization process resultsin the cutting of the TBS, using the inventive devices and processes,into viable morcelized tissue particles (MTPs) which are suspended inthe solution. Examples of MTPs include FTSGPs, split thickness skingraft particles (STSGPs) and cartilage particles (CPs), among others. Asstated throughout this specification, high viability of the MTPs isimportant to achieving a successful skin graft.

Stage 4 Separation/Dispensing

The MTPs in suspension are then filtered to remove the majority of thesolution, which generally increases the viscosity of the suspension.While some solution remains, most has been removed and the resultantmatrix of MTPs can be made to easily flow and spread on a wound ortreatment area. Depending on the mesh size openings of the filter, whichcan be chosen to fit the desired graft application and tissue type,small MTPs will be removed along with the filtrate. This portion ofsmaller particles in the filtrate may be used for other applicationssuch as injections for cosmetic use, or combined with additives such asanti-infectives or growth factors and used in the same application lateron in the further treatment of the patient. The filtered portion of theMTPs are then conveyed or pushed (for example with a plunger or withsuction) from the filter chamber into a dispenser (such as a removeablyconnected syringe), which is then used to apply or otherwise dispensethe MTPs onto the area of treatment to form the new skin graft.

Stage 5 Application to the Treatment Site

Upon completing processing with the device, the MTPs are received into afluidly coupled dispensing device, for example a dispensing syringe. Theprocess can be completed with minimal user handling of the excisedtissue. The matrix of MTPs in the dispenser is then ready forapplication to the treatment site. If the dispenser is a syringe orsimilar type dispenser, the matrix of processed MTPs can be readilyapplied by the physician or person administering the graft. The matrixof MTPs is intended to be easily spreadable and flowable, i.e.distributable, and the viscosity and density can be adjusted severalways, such as by determining the amount of solution to keep in thesuspension during filtering or by later adjusting the density of MTPsdistributed over the treatment site. Cellular outgrowth from the appliedMTPs will adjacently form new graft tissue, ultimately bridging betweenand interconnecting individual MTPs to cover the area of treatment.

The device of the present invention is configured to be employed with areusable, aseptically cleanable, operatable processor, used to drive themorcelizing mechanism within the device. Upon placing sterile saline andTBSs into the sterilized device, followed by activating the coupledoperatable processor, the MTPs are aseptically processed within thedevice and then received into a fluidly coupled dispensing device, forexample a syringe, ready for patient application. The viable tissue ismaintained within the aseptic system, suspended within sterile fluid,(i.e. saline or buffered saline) throughout processing. Processing iscompleted within several minutes and free of user handling.

The device has features which optimize processing cell viability andconvenience of tissue handling and transfer. Specifically, the aqueousprocessing allows for temperature, pH, and salinity control of theprocessing which would ideally be variable depending on the tissue,isotonic factors. The pH may be controlled with physiologic buffers. Thevariable blade speed allows for control of any potential baro-traumacaused by the formation of the vortex, which repeatedly moves the tissuesuspended in the aqueous medium through the cutting devices.

The selection of ultra-sharp cutting blades is one important factor inensuring that the tissue that is morcelized remains viable. The use ofpH controlled aqueous medium, along with the ability to control thetemperature of the medium during processing, as well as, the time, arealso important factors in achieving morcelization with the exceptionallyhigh degree of viability of the invention.

Mass-produced disposable razor blades and microtome blades are among thesharpest steel blades in the world. Razor type blades are typicallymartensitic stainless steel with a composition of chromium between 12and 14.5% and a carbon content of approximately 0.6%. The high-volumelinear process to produce such blades, starts with a roll formed stripof controlled thickness that is run as a ribbon through a continuousmanufacturing process. The linear manufacturing process enablesexceptionally tight and repeatable control of multiple sequencedautomated processes including, for example, grinding multiple distinctlystepped beveled/faceted cutting blade edges on both sides; with cuttingedges as thin as 30 nm for razor blades and 3 nm for microtome blades;with edges fortified with separate vacuum chamber applied hardenedcoatings (for example titanium+manmade diamond to harden edge), followedby, for example low friction polymer film for slipperier edge.Individual blades are progressively die-stamped in-line in a repeatablemanner.

Blade sharpness, is absolutely necessary to minimize cell mastication.Use of ultra-sharp blades, passing through and between masses of livecells, and through interstitial spaces, best assures a narrow margin ofviolated cells along the cut line with the least amount of shear forcesand crushing of cells. Slicing of tissue between two blade edges,passing at acute angles, enables stabilization of the tissue throughouteach cut to achieve controlled atraumatic slicing of whole thicknessskin tissue into morcelized particles.

Use of ultra-sharp blades, as achievable through automated processes,assures repeatability and expedites the morcelization process, enablingautologous whole thickness skin to be quickly converted into a newmorcelized implantable tissue particles within minutes.

To maintain cell viability, best practice is to keep the harvestedtissue wetted and then suspended in the pH controlled solutionthroughout handling (i.e. in saline, a buffer solution, or BioLifeSolution®, or other cell nurturing/preservation solutions, etc.).

Detailed Description of Devices

One preferred embodiment for morcelizing tissue, such as full thicknessskin grafts (FTSGs) and other tissues, supported in a fluid preferablysterile fluid, is shown generally in FIGS. 1-4.

Processing device 10 includes a liquid-tight container 12 having an openupper end 14 which may be suitably enclosed by a cover 16. The cover 16has an inlet aperture 18 which allows for insertion of tissue into thefluid. In a preferred embodiment the container is generally cylindricalhaving a closed curved bottom 20 opposite open upper end 14 with andexit opening 17 therein. While the cover 16 and the container 12 may bemade of various materials, in a preferred embodiment, the cover andcontainer are formed of a suitable plastic such as polypropylene (PP),polyethylene (PE), polystyrene (PS), polyethylene terephthalate (PET),polyimide (PA), acrylonitrile butadiene (ABS), polyetheretherketone(PEEK) and polyurethane (PU). Combinations or co-polymers of thesepolymers may be used. Glass, ceramic or metal containers may also beused. The container 12 may be transparent to visualize the processingand quality of the fluid and tissue being processed within.

An additional removable or adjoined protective cover that is able to bemanually opened and closed may be included so as to close off the coveropening during processing to best assure containment of fluid andcellular contents.

It is also contemplated that all components comprising the overallprocessing device (system), including the container and morcelizingmechanism, isolation device and applicator will be packaged and bulksterilized, for single-patient use and disposable. The packaged devicesmay be irradiated with gamma or e-beam or ethylene-oxide (EtO).Alternatively, the processing device may be sterilizable and reusable.Some portions of the system may be reusable and other portions may bedisposable.

Extending from bottom 20, container 12 includes a generally elongatetubular conduit 22 in fluid communication with the interior 13 ofcontainer 12 through opening 17. The conduit terminates in a containermount 24 at the lower end thereof. Extending outwardly and in fluidcommunication with conduit 22 is an outlet 26 which in the preferredembodiment shown in FIG. 1 extends at a right angle to conduit 22. Thedescription of the purpose of the conduit 22, the container mount 24 andoutlet 26 will described in further detail below.

Cover 16 is movably supported at the open upper end 14 of container 12for movement along a central axis A. The upper end 14 of container 12includes, for example, an outwardly directed key 12 a which is seated ina slot 16 a in adjacent skirt 16 b cover 16. The key 12 a is movablealong axis A within the slot 160 to allow for the movement of the cover16 with respect to the container 12, while restricting cover 16 rotationabout axis A.

The cover 16 further includes an inwardly formed downwardly extendinggenerally tubular stem 28 having an upper cup-shape cavity 30 covered bya cap 31. The stem 28 accommodates a mounting rod 32 having a threadedlower end 34 and an upper end 36 terminating in an enlarged head 38. Thehead 38 is captively retained within the cavity 30 supported by a spring39, for example by one or more Belleville, dome, single or multiple wavetype washers, captive between the lower end of cavity 30 and theenlarged head 38. The spring or springs 39 may additionally be captivelysandwiched between conventional type washers 39 b, the threaded lowerend 34 of rod 32 is threaded into impeller 108 about axis A. A shoulder32 b, adjacent to threaded end 34 of rod 32, is supported againstimpeller 108.

A disk shaped stationary cutting member 102 is supported upon theterminus of stem 28 on a perpendicular plain relative to axis A. Thestationary cutting member 102 is constrained from rotating about axis Aby, for example, mating pins 28 a or other keyed features in engagementbetween the stem 28 and the stationary cutting member 102.

Now as best shown by FIGS. 1-4 and FIGS. 9-10, the sub-assembled rod 32,head 38, impeller 108, and blades 112 are together captured andconfigured to rotate about axis A relative to stem 28. The spring orsprings 39, between head 38 and the lower end of cavity 30, act to lifthead 38 and thereby lift stem 28 and impeller 108 through stem 38 tocontinuously constrain rotating blade edges 112 a in compression againststationary cutting member 102.

Referring particularly to FIGS. 5-11B, the subassembly of rotatingimpeller 108 with associated rotating blades 112, held compressivelyagainst stationary cutting member 102, about axis A by means of a rod 32with head 38 and lower end 34 and spring 39 through stem 28 arecollectively referred to as a morcelizing mechanism.

As described in further detail below the drive shaft 42 is attached toan operatable processor 44 shown schematically in FIG. 1 by way of adrive engagement 46 at the lower end of drive shaft 42. The processor 44causes rotation of the drive shaft 42 and thereby rotation of therotating blades 112 against the stationary blade edges 122 within themorcelizing mechanism 40 which causes morcelization of the tissue, orspecifically FTSGs within in the container 12. A suitable rotary shaftseals 47 and 41 provide a fluid seal between drive engagement 46 anddrive shaft 42.

The processor 44 is preferably a reusable device that is configured forease of being aseptically cleaned following each use. The processor 44is also configured to replaceably receive a processing device 10 inmechanical engagement in such manner that enables a user to attach, use,and remove the processing device using standard practices forinterfacing surgical devices in a sterile field. The processor 44 mayalso be configured to receive a sterile drape to isolate surfaces notshrouded by a coupled processing device. The processor may also be adevice that is entirely sterilizable and/or powered by compressed airthat is easily available in the operation room setting.

As shown in FIG. 2, and described in more detail below, rotation of theimpeller 108 provides a continuous circulating flow (CF) of the fluidand contained tissue or specifically FTSGs about the interior ofcontainer 12 and through the morcelizing mechanism 40 so as tocontinually cut the FTSGs into progressively smaller particles. Themorcelizing mechanism 40 is seated in fluid-tight relationship over exitopening 17 in the open bottom 20 of container 12 to maintain the FTSGsand fluid within the interior of container 12 throughout morcelization.A suitable seal 19 provides a fluid seal between morcelizing mechanism40 and exit opening 17.

As is shown in FIGS. 3 and 4, the drive shaft 42 may be raised so as tounseat the morcelizing mechanism 40 from opening 17 in the open bottom20 of container 12. Upward movement of drive shaft 42 along axis Acauses upward movement of the cover 16 with respect to the container 12with the key 12 a riding within slot 16 a. This lifts morcelizingmechanism 40 off of its sealed position on the bottom 20 of container 12thereby rendering accessible exit opening 17 for fluid flow.

Morcelizing Mechanism

Operative components of morcelizing mechanism 40 are shown in furtherdetail with additional reference to FIGS. 5-8.

Morcelizing mechanism 40 includes a base component 100 and a stationarycutting member 102 which are axially aligned over one another along axisA. Base component 100 is mounted to the drive shaft 42 with a dependingmount 104 to provide for rotation. Above mount 104 is a flat circularplate 106 which is generally transverse. Plate 106 also serves as theseating surface in opening 17 of the bottom 20 of container 12 as isshown in FIGS. 1-4.

The upper end of base component 100 serves as an impeller 108 having twoor more impeller vanes 110 upwardly extending from plate 106 ondiametrically opposed sides of axis A. The impeller vanes 110 are eachcurved along axis A in a complimentary fashion for purposes that will bedescribed in further detail below. Each impeller vane 110 supports infacing relationship at the upper end a cutting blade 112. As also willbe described in further detail below, the cutting blades 112 at theupper ends of impeller vanes 110 are supported in juxtaposition with thestationary cutting member 102. The blades 112 rotate with base component100 with respect to stationary cutting member 102.

In a preferred embodiment shown in FIGS. 5-8, the stationary cuttingmember 102 has generally disc shaped body 102 a. The body 102 a definesspaced apart blade surfaces 120 arranged circumferentially. Each bladesurface 120 includes a pair of converging blade edges 122 which convergeat an apex 122 a. In between each of the blade surfaces, breaches 124are defined. The breaches 124 are open spaces between the blade surfacewhich permits passage of the TPs, such as FTSGs and other TPs, and fluidthrough body 102 a as the base component 100 rotates.

In one embodiment shown in FIGS. 7 and 8, the stationary blade edges 122are defined by longitudinal radially extending members 123 convergingwith an arc of the circle forming the outer edge of the disc shaped body102 a.

In a more preferred embodiment shown in FIGS. 5 and 6, the bladesurfaces 120 are formed in a tear drop shape where the apex 122 a of theconverging blade surfaces 120 converge near the circumference of thestationary cutting member 102 in a tapered curved surface. It has beenfound that this shape helps promote complete morcelization of the tissueand specifically TPs passed through.

The arrangement of the stationary cutting member 102 with respect to theimpeller 108 is shown schematically in FIGS. 9-12. A small clearancespace (S) is provided between the lower edge of the stationary cuttingmember 102 and the upper end of impeller 108 such that the extendingrotating cutting blades 112 are supported in juxtaposition against thelower edge of the disc shaped body 102 a of stationary cutting member102. This creates a shear plane (SP) at which the tissue is sheared andmorcelized.

The stationary cutting member 102 is preferably stainless steel and CNCmachined with precision ground sharp burr free stationary blade edges122. The bottom shearing plane (SP) surface must be flat and preferably0.08 μm or better finish. The stainless steel material may generally bea corrosion resistant and hardened grade, for example 440C stainlesssteel, machined in annealed state and vacuum heat treated to 55-60 RC toachieve a hardened surface and durable sustainable cutting edges. Thestationary cutting member 102 may alternatively be of othernon-corrosive materials or may be of an alternative hardness and may bemade by other precision process. The stationary cutting member 102 andjuxtaposed rotating cutting blades 112, in compressive engagement, maybe of differing materials, such as, for example, plastic or ceramic, orof differing hardness, or have alternative treated, or applied surfacefinishes, to best avoid wear or galling conditions as opposing surfacesslide upon each other along a shear plane (SP). Additionally, the bladeedges 122 of the stationary cutting member 102 and/or the blade edges112 a of the rotating cutting blades 112 may be formed to be sharp orsubsequently sharpened.

The rotating cutting blades 112 may be mounted at the upper end of theimpeller 108 supported by a spring such as an elastomeric pad 130 whichbiases the edge of the cutting blade 112 against the lower edge of thedisc shaped body 102 a of stationary cutting member 102. It has beenfound that maintaining the cutting blade edges 112 a in physical contactagainst the stationary cutting member 102, minimizes tearing andshredding of the tissue.

Referring to FIGS. 5 and 6, the stationary cutting member 102 may havetwo or more breaches 124 radially arrayed about axis A, preferably asshown three, each with associated blade edges 122. The rotating basecomponent 100 may similarly have two or more cutting blades 112 radiallyarrayed about axis A, preferably as shown two. However, the quantity ofrotating blades 112 best differ from the quantity of stationary blades122, so as to maximize cutting efficiency by minimizing otherwisecumulative cutting forces as would be compounded should multiple bladesengage simultaneously.

The rotating cutting blades 112 are positioned at an acute cutting anglerelative to the juxtaposed stationary blade edges 122, such that tissue,when captured between the rotating blades 112 and juxtaposed stationaryblades 122 will be cut with a slicing action.

FIG. 12A shows that cutting edges 112 a of the rotating blades 112 maybe manufactured with a ground double beveled edge. Double bevel refersto beveled on both sides of the blade. Alternatively, as shown in FIGS.12B and 12C, the cutting edges 112 a can be made sharper with asecondary distal honed double beveled edge which further maximizes themorcelization of the TFSGs. Honing refers to a more precise abrasivegrinding or lapping process in which a relatively smaller amount ofmaterial is removed from the surface by means of a finer grit abrasive.The cutting blades 112 used in our functional proof-of-principle systemsutilize preferably further sharpened blades which have a secondary honeddouble beveled edge, as well as an additional finely honed doublebeveled edge, for example three graduated sets of double beveled edges.

The rotating cutting blade 112 is best arranged at an acute anglerelative to the lower surface of the stationary cutting member 102, sothat the tip of rotating blade cutting edge 112 a passes at an acuteangle with respect to the stationary blade edges 122 of blade surfaces120.

As previously described above, springs 39 may be used to compressivelypre-load the rotating blade edges 112 a to maintain contact uponstationary blade edges 122 throughout rotation. FIG. 12C shows that thecutting edge 112 a of the rotating blades may flexibly conform underpreload against the shear plane (SP), particularly when the cuttingblade 112 may be substantially stiff, for example approximately 0.010inch thick. Additionally or alternatively, as shown in FIG. 9, thecutting blade 112, itself, may flexibly conform under preload againstthe shear plane (SP), particularly when the cutting blade 112 may besubstantially flexible, for example approximately 0.003 to 0.006 inchthick.

FIG. 12D shows an embodiment where the blade edges 112 a of the rotatingblades 112 may be formed to have a flat or planar extent 112 b. Thisplanar extent 112 b is formed to be co-planar or co-extensive with theblade edges 122 of the stationary cutting member 102 at shear plane(SP).

While in a preferred embodiment, the shear plane (SP) is normal to thechamber impeller and blade rotation access. The shear plane (SP) mayalso take other direction with respect to the axis A. One example isshown in FIG. 13A which shows a bushing 200, a rotatory seal 210, astator 220, a rotor 240 and a cutting chamber 260. The blades 242 ofrotor 240 pass in close proximity to the blades 222 of stator 220 whichare stationary blades about a central axis A.

Also, in this embodiment, the rotor blades 242 have surfaces which areconfigured as integrally formed impeller vanes. The rotating edges maybe generally co-extensive to the leading impeller vane edges. Therotating rotor blades and stationary stator blade edges shouldpreferably remain in intimate physical contact to best achieve preciseslicing. The blades may be machine honed for closely controlled minimumshear gap, preferably less than 30 micrometers. Positioning the rotatingblade edges at an acute angle relative to the shear plane of stationarycutting blade edges facilitates a shear cut for the impinged tissue.Maintaining a spring assisted compressive engagement between therotating blade edges and the shear plane of the opposing blade edgesbest assures that tissue will be precisely slice, rather than to slipbetween the converging blade edges.

Other techniques and arrangements for cutting the tissue at a shearplane may also be within the contemplation of one skilled in the art.

Tissue Morcelization

Having described the basic components of the process device 10 of thepresent invention, one preferred example of the morcelization of tissueinto particles (MTPs) or specifically tissue particle (TPs) as definedherein will be described with respect to the Figures. The term “tissueparticles” is also refereed to herein as TPs.

Initially, with reference to FIGS. 1-4, FTSGs prepared as abovedescribed and in a fluid medium may be inserted into container 12through inlet aperture 18 of cover 16. A fill line F may be provided soto provide guidance as to the volume of tissue and fluid which may beplaced in container 12. Thereafter, the rotating mechanism connected todrive engagement 46 is activated so as to cause rotation of drive shaft42 and morcelizing mechanism 40.

Referring to FIG. 2, such rotation causes circulating flow of the tissuein the fluid by establishing a vortex within container 12. This vortexprovides for continually moving the tissue through the morcelizingmechanism so as to fully morcelize the tissue contained therein. Thecirculating flow path as well as the vortex established is created bythe configuration of the impeller vanes 110 of the impeller 108.

Shown schematically in FIG. 14, the impeller vanes 110 are constructedso that an upper or leading portion 110 a of the impeller vane 110imparts an axial thrust upon the fluid and contained tissue (i.e., TBS)or specifically FTSGs, while a lower or terminal portion 110 b of theimpeller vane 110 provides for radial thrust. The construction of theimpeller vanes 110 a and 110 b provide for continually moving the FTSGsthroughout the container 12. The impeller 108 causes fluid withcontained FTSGPs to be driven through breaches 124 in the stationarycutting member 102, passing between rotating blades 112 and stationaryblades 122, to then be deflected against the trough-like bottom 20 andside walls of container 12 to reverse the flow in the oppositedirection, circulating through the outer peripheral volume (PV). Thefluid flow then transitions into a vortex to mix and drive the tissuethrough the central volume (CV) to return again through the morcelizingmechanism 40 so that the FTSGs are continually and repeatedly cut andmorcelized.

A person of ordinary skill in the art will be able to alternativelyconfigure, for example, impeller vanes and/or internal containergeometry and/or bottom 20 forms so as to enhance effective circulatingflow of fluid with suspended tissue though-out the container 12 andthrough the morcelizing mechanism 40. Internal flow characteristics maybe enhanced, for example, by increasing or decreasing the pitch orotherwise reshaping the form of impeller vanes 110; or by increasing ordecreasing the pitch of a portion of a vane configured for axial thrust110 a relative to the pitch of a portion of a vane configured for radialthrust 110 b; or to eliminate either of the axial thrusting vanesurfaces or radial thrusting vane surfaces.

Referring to FIGS. 3 and 4, once the TPs, such as FTSGs and other tissueparticles as described herein, are fully morcelized, the drive shaft 42may be raised, unseating the impeller 108 from its seated position inthe container. The plate 106 is unseated from opening 17 establishingfluid communication with conduit 22 and outlet 26. The morcelized tissueis discharged by a gravity driven drain through outlet 26 for use in amanner which will be described hereinbelow.

Upon completing the morcelization of FTSG or other tissues grafts intoMTPs, the impeller 108 rotation may be stopped and the MTPs, as definedherein, having a specific gravity greater than water, will settle to thebottom 20 of container 12. One skilled in the art will recognize thatalternative methods may be used to manually withdraw the settled MTPsfrom the bottom 20 of container 12 within the processing device 10. Forexample, the settled MTPs may be drawn into a conventional syringe incombination with an elongated cannulated tip (not shown) that can beinserted into the container 12 through the inlet aperture 18. In suchmanner, the same syringe used to draw the MTPs from the processor 10could then be used as an application device. In this manner of manuallydrawing out the MTPs through the inlet aperture 18, the processingdevice 10 need not include a drain 39 or an outlet 26.

Discharge of Morcelized FTSGPs

Discharging the morcelized Tissue Particles (MTPs) or for examplespecifically FTSGPs, into an applicator 300 may now be described withrespect to FIGS. 15A-E. Referring to FIG. 15A, an applicator 300, whichmay be used to collect and dispense the MTPs.

TPs is typically configured as a syringe which provides a well-knownmeans to deliver and meter out controlled volumes.

The syringe applicator 300 also serves as a device to separate excessfluids from the MTPs. A cylindrical screen filter 302 is formed as aninsert to the applicator body 303 and has an inner applicator chamber308 lumen to receive a plunger 304 and piston 306. Peripheral drainchannels 305 may surround the filter 302 such that excess fluid withinthe MTPs may pass freely through the filter walls, through the drainchannels 305 and out the drain outlets 310. The screen filter mayencompass 360° of the applicator or only a portion thereof, as shown inFIG. 15A, so as to leave sufficient area to view the contents through awindow 307. The filter 302 may be a fine mesh, for example having 50micron openings allowing fluid to pass through while containing MTPs.Alternatively, filter 302, may be comprised of, for example, laser cut,woven mesh or acid etched perforated screens with specifically sizedlarger openings, so as to drain away smaller particles with solution,while selectively containing wetted particles larger than the utilizedfilter openings. The syringe applicator 300 could also be configured toseparate liquid from the MTPs and dispense the MTPs onto or into aseparate device or container for application, for example, into anattached syringe.

FIG. 15A also shows that the processing device 10 is attached onto theprocessor 44, for example, with a bayonet mount 115. The processingdevice 44 may be cordless and include a low voltage DC motor 114 drivenby a contained rechargeable battery. The battery is preferably rechargedby connection to a remote ACDC charger. The low voltage DC motor mayalso be powered through the remotely connected ACDC power source. Inboth such manners the use of low voltage DC power enables the safe useof processing device 10 in the potential presence of an aqueoussolution. The axially connected drive engagement 46 connects the motorshaft of the processor 44 to the central shaft assembly. Uponmorcelization of the TPs, the motor 114 may automatically be slowed orstopped and raised so as to open the seal 19 below the impeller, causingthe morcelized tissue mixture and solution to drain through the gravitydriven drain 39 from the processing chamber through the outlet 26 andthrough the port 320 to enter the applicator 300.

The applicator 300 is shown with the plunger 304 and piston 306 in itsraised position and with the cap 318 in place to close the dispensingorifice. As the MTPs and solution enters the applicator 300, the fluidis drained away from the MTPs as the fluid will freely pass through thefilter walls 302, through the flow channels 305 and exit the applicator300 through the drain outlet 310 into the fluid waste drain container317. Thereafter, the plunger 304 may be advanced sufficiently into theapplicator chamber 308 so as to enable the piston 306 to close off theport 320. The applicator 300 may then be removed from the processingdevice 10 by disconnecting inlet port 320 of the applicator 300 from theoutlet port 26 a of the processing device 10. Thereafter, the cap 318may be removed from the applicator 300 and a selected applicator tip 301may be affixed to the dispensing orifice 309 for dispensing the MTPs ina manner which will be described in further detail hereinbelow.

A further embodiment of the present invention, shown in FIG. 16, issimilar to FIG. 15A relative to including a processing device 10mechanically coupled onto a processor 44. The drive engagement 46 maysimilarly be raised to open seal 19 to drain the container 12 through anoutlet 26, however, as shown in FIG. 16 (as well as in various earlierFIGS. 1,2,3) the drive shaft 42 may include a rotor pump 118, forexample with fins integrally molded upon the drive shaft 42. The rotorpump 118 is driven by the motor 114 through axis A, to circulatesolution through an isolation device 415.

FIG. 16 introduces a different isolation device 415 which circulatesfluid from the processing device 10 through outlet 26 and inlet 29conduits. Upon completing morcelization of the TPs, the motor 114 willbe automatically raised, along with drive engagement 46 and drive shaft42, so as to open the chamber seal 19 below the impeller 108, to releasefluid and MTPs from container 12.

The motor speed is changed, as appropriate, to pump the solution and TPsthrough the outlet channel 26 and through a diverter valve 417 to entera cylindrical isolation chamber 416 containing a cylindrical filter tube402 lining. The filter tube 402 may be, for example a woven mesh,perforated film or acid-etched screen with openings sized appropriatelyto capture particularly desired sized MTPs. The particles are capturedwithin the isolation chamber 416 as fluid passes through the filtertube, and through circumferential drain channels 405, exiting theisolation device 415 through inlet conduit 29, to return into thecontainer 12 of processing device 10. Within several brief passes themotor 114 will automatically stop as the MTPs are substantially rinsedaway from the container 12 and transferred into the isolation device415.

The diverter valve 417 may then be automatically or manually switched,for example turned 90° clockwise, to open a fluid path from theisolation chamber 416 into the applicator 400. The inlet conduit 29 ispositioned below the fill line (F) of container 12, enabling the headpressure of contained fluid to substantially flush the MTPs from theisolation chamber 416 through the diverter valve 417, through anapplicator attachment 411, and into the applicator chamber 408 of adetachable applicator 400.

Thereafter, the diverter valve 415 is closed; the applicator 400 isdisconnected from the applicator attachment 411; and a plunger withpiston (as shown for example in previous FIG. 15B) is manually insertedinto the lumen of applicator 400. The cap 418 is removed (for examplewith a Luer type connection) and replaced with a selected applicator tip(as, for example introduced in FIG. 15B). In this manner, the applicator400 is ready to dispense the MTPs matrix to a desired autologous implantsight in a manner which will be described in further detail hereinbelow.

The processing device 10, isolation device 415 and applicator 400 may bepackaged as an integral sterile assembly. Applicators 400 mayalternatively be sterile packaged separately.

Turning now to FIG. 17, a further embodiment of the processing device 10with processor 44 and applicator 500 is shown here coupled to adifferent type of isolation device 515. In this embodiment a pump 118will similarly circulate the solution with suspended MTPs from thecontainer 12, through outlet 26 and returning through inlet 29 both influid communication with an isolation chamber 515 which in thisconfiguration employs cyclonic action to separate the MTPs from thesolution. The system uses the principle of terminal settling velocity ofsolid particles in a centrifuge field. The outlet 26, from theprocessing device 10, enters tangentially into the isolation chamber 516of the isolation device 515. High velocity centrifuge fields within thehydro cyclone cause particles to migrate rapidly to the outside walls ofthe conical chamber 516 and will be forced to move downward on theinside of the conical walls through a valve 517, through an applicatorattachment 511, and into the applicator 500. A valve 517 may then beclosed, the applicator 500 is disconnected from the collector, a plungerwith piston is inserted into the applicator 500, the applicator cap 518is replaced with the dispensing tip of choice, whereupon the applicatoris ready to dispense MTPs in a manner described hereinbelow.

A still further embodiment is shown in FIG. 18 where the processingdevice 10 is coupled to the processor 44. In FIG. 18, the inlet 20 andoutlet channels 26 are shown in fluid connection with an isolationchamber 616 that employs a whirlpool like action to gather the swirlingMTPs particles towards the central drain through which the concentratedMTPs will be deposited into an applicator chamber 608 within anapplicator 600. As with the above embodiments, the valve 617 is thenclosed, the applicator 600 is disconnected and the applicator cap 618 isreplaced with a dispensing tip of choice. The applicator 600 is thenready to dispense the MTPs in a manner which will be described infurther detail hereinbelow.

A still further embodiment is shown in FIG. 19 where the processingdevice 10 is coupled similarly as shown in FIG. 16 onto an isolationdevice 716 through an outlet 26 and an inlet 29. Also similar to theembodiment of FIG. 16, the isolation device 716 contains an isolationchamber 716, separated by a filter tube 702 from a drain channel orchannels 705, such that solution passing through the isolation chamber716 will pass through the filter tube 702, to pass through the drainchannel 705, to pass through the inlet 29 and be recirculated throughthe processing device 10. However, in this embodiment, the outlet 26 andinlet 29 may include sealable closable ports 720 (not shown) such thatthe isolation device 716 may be detachable from the processing device 10while containing fluid from leaking from the detachable outlet 26 andinlet 29 flow paths. In this manner the detached isolation device 716may contain a plunger 704 and piston 706 and detachable cap 718 andtogether may be used as applicator 700 as similarly described in FIGS.15C and D.

The schematically drawn circulating flow (CF) paths in FIGS. 2 and 14have been significantly simplified, by not indicating the turbulentvortex swirl, so as to more clearly depict the recirculating nature ofthe fluid flow pattern. Early prototypes revealed that a vortex inducedby the impeller, while desirable to continuously recirculate and mix thefluid suspended TPs, also caused the particles to travel many morecircuitous times around the container 12 than necessary before beingdrawn through the morcelizing mechanism 40 and becoming morcelized.

FIG. 20, an axial view, and FIG. 21, a lateral view, introduce apreferred improvements to the processing device 10 to more expedientlymorcelize TPs. Placement of vertical baffle panels 140, radiating fromthe stem 28, effectively interrupt the single vortex. The baffles arepositioned proximal to the apex of each set of converging blade edges122 on the stationary cutting member 102. The swirling fluid within thecontainer 12 rebounds off each baffle 140, creating a separate smallervortex V adjacent to each baffle 140. The smaller vortexes V carry theTPs (tissue particles) more expediently through each of the continuouslyclosing breaches 124 of the morcelizing mechanism 40.

Although the circulating flow (CF) paths within processing device 10 asdescribed in FIGS. 2 and 14 represents a preferred embodiment (and isalso included by way of example in multiple other Figs.), it is notintended to limit the scope of the invention. Whereas an impeller 108 isemployed to continuously recirculate fluid and TPs through the centralvolume (CV) of container 12 so as to repeatedly pass through themorcelizing mechanism, the fluid and TPs need not necessarily berecirculated through the peripheral volume PV upon return. FIG. 22,therefore, teaches that the fluid and suspended TPs may be recirculatedthrough the morcelizing mechanism 40 in other manners, by way of anotherexample, to flow externally of the container 12, through a recirculatingconduit 150.

Further, whereas FIG. 22 shows a rotor pump 118, integral to shaft 42and rotating about axis A, one skilled in the art would recognize that afluid driving pump may alternatively be included elsewhere along arecirculating conduit 150 between an outlet 26 from the processingdevice 10 and a return inlet 29 to the processing device 10. Further, assuch, a circulating pump (not shown) need not be driven by or associatedwith a motor also used to drive the processor 10 and could be, forexample, a separately operable fluid pump. Further, referring still toFIG. 22, such a recirculating conduit 150 may include one or morediverter valves 117, such that (upon completion of morcelization) thefluid and suspended MTPs can be diverted to circulate through any ofvarious types of isolation devices, for example as described throughFIGS. 15 A-E, 16, 17, 18 or 19.

Further still, the impeller 108 of FIG. 22, used in a system configuredwith a recirculating conduit 150, external to the container 12 ofprocessing device 10, need not have vanes configured for radial thrust110 b. In such an embodiment, vanes with a pitch configured for axialthrust 110 a alone may be sufficient on an impeller 108 to facilitatecirculation of the tissue bearing solution from container 12 along axisA, through outlet channel 26, through a recirculating conduit 150, andthrough inlet channel 29 to be continuously recirculated through themorcelizing mechanism 40 within container 12.

A still further embodiment is schematically shown in FIG. 23. Here aprocessing device 910 is shown used in conjunction with a centrifugaltype of isolation device 919 to effectively separate and compactprocessed MTPs, as defined herein, for dispensing through a conventionalsyringe (not shown). The isolation device 919 may be shrouded within aprotective cover 976. The processing device 910 and isolation device919, together with enclosing shroud 976, may preferably be integratedinto a single unit, to be packaged and pre-sterilized as a singlepatient use as a disposable device as red bag medical waste. The devicemay be used multiple times within a procedure for an individual patient.The combined processing device 910 and isolation device 919 areconfigured to be axially aligned and fixably coupled, for example with abayonet engagement 960, onto an aseptically cleanable reusable processor944.

Upon completing the morcelization of TPs within a processing device 910,the MTPs are released in solution through a drain 939 from the bottom920 of container 912. The drain 939 is preferably located about acentral axis A of the processing device 910 or otherwise appropriatelylocated on the bottom 920 of the container 912, so as to fluidlycommunicate into a central chamber 972 of the centrifugal isolationdevice 919.

One or more individual collection chambers 973 protrude radially fromthe central chamber 972, each collection chamber having a distal outletorifice 974. The distal outlet orifice 974 has a standard threadedfemale Luer engagement for interchangeable attachment of a standard Luercap 975 or a standard Luer tipped syringe (not shown).

The central chamber 972 and radially extending collection chambers 973may be integrally formed as a hollow injection blow molded component, orproduced as an assembly of injection molded components, or acombination, for example with injection molded Luer fittings affixedonto an injection blow molded unibody core.

The centrifugal isolation device 919 is configured to rotate at a highspeed, for example up to 300 Gs, on precision radial type ball bearings(not shown), about an axis that is preferably coincident to or axiallyaligned with axis A of the processing device 910. The spinning isolationdevice 919 is preferably encased within a protective shroud 976. Shouldthe Processing device 910 and the centrifugal isolation device 919rotate about the same axis, the ball-bearing's inner shaft diameter maybe sized sufficiently large as to enable independent rotation of thecentrifugal isolation device 919 relative to rotation of the impeller908 within the processing device 910.

Upon being centrifugally spun for only a few minutes, the solutionsuspended MTP's will separate and become compacted within the radiallyextending collection chambers. The Luer caps 975 on the distallyextending female Luer connectors 974 are then unthreaded and exchangedwith appropriately sized standard syringes. Upon then drawing thecompacted MTPs into the syringes, the filled syringes are disengagedfrom the isolation device 919 and a selected Luer fitting applicator tip(for example as previously described in FIG. 15) is affixed, now readyfor autologous MTP application to the intended site.

Morcelized Particulates

A solution of suspended MTPs, e.g., FTSGPs or other tissues particles asdescribed herein, may be mixed in combination with other FDA approvedadditives, for example handling (i.e. in saline, a buffer solution, orBioLife Solution®, or other cell nurturing/preservation solutions,etc.). The mixture may be created within the processing device 10, usingthe vortex circulation to achieve a heterogenous mixture of morcelscomprised of naturally connected cellular and extracellular matrixmaterial. Alternatively, some suspensions/dispersions of MTPs may behomogeneous. Whether the dispersion or emulsion produced is homogeneousor heterogeneous may depend on a number of factors, including withoutlimitation, the type of tissue(s), the medium it is suspended in, thespeed and temperature of the process, among other factors. An importantadvantage of the method of creating the MTPs suspension/dispersion ofthe invention, as well as the resultant suspension/disperisons per se,relates to the high cellular viability during and immediately afterprocessing to achieve the morcelization. This ability to morcelizedwhile maintaining such high cellular viability, as described herein, isunique to the present invention and not achieved by prior methods. Themethods described herein produce suspensions or dispersions whichcontain MTPs having at least 50% viability immediately after processing,which is generally in real time at the bedside of the patient; or atleast 60% viability immediately after processing; or at least 70%viability immediately after processing; or at least 80% viabilityimmediately after processing; or at least 85% viability immediatelyafter processing; or at least 90% viability immediately afterprocessing; or at least 92% viability immediately after processing; orat least 94% viability immediately after processing; or at least 96%viability immediately after processing; or at least 97% viabilityimmediately after processing; or at least 98% viability immediatelyafter processing; or at least 99% viability immediately afterprocessing. Generally, the processing may take about 1 hour, butdesirably less than 1 hour, for example, 45 minutes or less, 40 minutesor less, 30 minutes or less, 20 minutes or less, or 10 minutes or less.

The MTPs may also be centrifuged to vary the density, viscosity andconsistency of the tissue particles, as may be desirable for alternativesurgical applications. Modulating the centrifuge speed and duration ofcentrifuging enables the customization of the resultant output tissueparticle form, for example the consistency and density may present as asolution, or a paste, or a cream. Desirably, the resultant output (Id.)flowable and/or easily applied by spreading. The resultant output mayfurther be presented, for example, as compacted tissue form or may befurther spun to present as compacted cellular matter.

The MTPs as defined herein may be most efficiently delivered from avariety of fluid dispensing devices, most notably syringes, which arefamiliar and useful to easily meter controlled volumes. The targetedparticulate sizes will pass freely as a fluid composition through thelumen of standard Luer connectors. Tips may be interchangeably attachedonto an applicator, for example, with a standard Luer thread. A varietyof interchangeable applicator tips may be included within a dispensingkit for selection as most appropriate for a specific application at theoption of the surgeon for a given procedure. In a cream, paste or fluidform, the MTPs, for example FTSGPs, may be dispensed from a syringethrough various selected tip types of applicator tips. A tip may have anarrow/long fanned outlet orifice to spread over a large area. Such afanned tip may be comprised of a flexible low durometer silicone orthermoplastic elastomer and may have a thin flexible edge, so as to beuseful to gently and evenly spread the MTPs over large and/or irregularwound surfaces, for example burns.

In a dense form, the MTPs of the invention may be spread or applied overareas or into crevices, for example with a spatula. In a dense form, theMTPs, for example Cartilage Particles (CPs), may be used as a filler,for example for cartilage defects. As such the MTPs, such as CPs orother morcelized tissues particles as defined herein, may be mixed withfibrin glue, autologous Platelet Rich Plasma, growth factors or otherFDA approvable materials, for example as a binder. Cartilage or organMTP's may also be delivered through an endoscopic syringe attachment.

In a fluid a cream or solution form partial thickness dermal skin graftparticles may alternatively be dispensed from a syringe, through aflexible cannula or a needle, for subdermal applications, for example,to fill cosmetic defects. For delivering the various MTPs as describedthe lumen may range, for example, from 22 to 18 gauge, or most notably22 or 21 gauge.

In a highly soluble or dispersible form the MTPs, which in such a casemay be of the smaller variety, as described herein, may be sprayed overlarge areas. In the case of MTPs for use on burns or open wounds, anon-adherent surgical wound dressing, may be used to prevent the appliedMTPs from migrating while keeping the wound site moist and protectedfrom infection. Such commercially available dressings include, forexample, commercially available Drawtex, Sofsorb, Kalginate, or Aquasorbdressings.

System Methodology

A system of devices is described to perform a process in a methodicalsequence. With reference additionally to FIG. 24, which shows the systemprogression in operation, the system and process includes: 1) aprocessing device 950 is used to introduce and morcelize tissue intotissue particles (MTPs) within a solution; 2) an isolation device isthen used to separate for dispensing morcelized tissue particles (MTPs)suspended in solution from the preponderance of solution; and 3) anapplicator device 952 is then used to surgically dispense and apply thecollected morcelized tissue particle matrix. Heretofore we havedescribed several alternatively configured devices and methods employingnon-limiting details with which to accomplish a three-step process toachieve the desired outcomes.

A processor device may be used in conjunction with various types ofisolation devices. For example, an isolation device may take the form ofa device including a filter tube through which solution flows to isolateMTPs as described in FIGS. 15 A-E, 16 and 19; or an isolation deviceutilizing cyclonic action within a chamber to isolate MTPs as describedin FIG. 17; or a device using a whirlpool action within a chamber asdescribed in FIG. 18; or a device performing as a centrifuge asdescribed in FIG. 23; or the MTPs may simply be isolated by sifting thepreponderance of solution away through a screen (not shown); or settledMTPs may be drawn from the solution using a standard syringe; or anynumber of other methods may be contemplated to isolate MTPs in suspendedfrom the preponderance of the solution.

A processor device may also be used in conjunction with various types ofapplicator devices. For example, an applicator device may be a standardsyringe; or an isolation device may additionally be deployed for use asan applicator as described in FIG. 15A-E or 19: or a spatula may be usedto manually apply the MTPs matrix; or the tissue particulates in liquidsuspension may be sprayed over large wound areas; or any number of othermethods may be employed to deliver and apply MTPs in a controlledmanner.

FTSG Process Verification Studies

Full Thickness Skin Grafts (FTSGs) harvested from a human abdominoplastywere prepared in accordance with the methods disclosed herein and usingthe apparatus and systems disclosed herein.

Harvested sample FTSGs of various noted sizes were each separately andindividually placed into fabricated experimental test apparatusesmodeled as generally described in FIGS. 1-4 without baffles and FIGS.20-21, with baffles. The processing devices were filled with 35 ml ofbuffered saline solution, pre-chilled with ice chips. The FTSGs werethen morcelized by subjecting the samples to a slicing speed ofapproximately 550 rpm for incrementally stepped durations, timed inminutes at ambient room temperature of 70° F. Earlier tests demonstratedinsignificant temperature rise of the chilled buffered saline water overthe lapse time processing each sample.

MTPs were quantitatively assessed to determine cell viability. ProcessedFTSGPs suspended in solution were transferred using a syringe into 0.5ml aliquots. The aliquots were maintained chilled in a container ofchipped ice. The samples were spun down in a centrifuge at 700 RPM forfive minutes followed by removing the supernatant. Quantitative cellviability analysis was performed using standard trypan blue testprotocols to stain and count living cells versus purple ruptured deadcells. A table included as FIG. 25 documents highly viable cellularviability test results ranged consistently between 87% to 98%viability—across multiple exemplary sample lots and processingparameters and processing durations, a sampling of which are describedbelow.

Additional MTT tests yielded similar quantitative results toconsistently confirm data reliability. MTPs were also qualitativelyassessed. Resultant morcelized tissue particles, suspended in solution,were drawn from the processor through a cannula into a syringe andexpelled into a petri dish to form a shallow pool or puddle aside ametric scale. Photos of the morcelized FTSGPs are shown, herein, eachphoto identified by test sample numbers, to visually and qualitativelydocument relative morcel sizes and particle appearance. As evident inthe images, tissue particle sizes are relatively consistent within eachsample. Maximum particle sizes became progressively smaller with longertotal duration of processing time.

In exemplary morcelization studies, ten portions of full tissue skingrafts (FTSGs), each approximately 12 mm×6 mm×4 mm thick were morcelizedin a processor without baffles, containing 35 ml of buffer solution withthe blades rotating at approximately 550 RPM. FIG. 26A (test 1b) shows asample of morcelized FTSGPs in fluid suspension drawn from the processorafter 4 minutes of processing. FTSGPs in solution were transferred toform a shallow pool in a petri dish to visualize individual particles.The maximum sizes of individual particles appear to generally be no morethan approximately 1.5 to 2.0 mm on any axis. The majority of particlesizes appear less than 1 mm FIG. 26 shows a subsequent sample then drawnafter an addition three minutes, for a total of 7 minutes. The secondsample appears more densely populated with particles and the typicalmaximum particle sizes appears to have been reduced to no more than 1.5mm across.

In a next exemplary morcelization study, still using the same patienttissue, the processing chamber included three baffles and four (versus10) portions of FTSGs. The sample was again processed with 35 ml bufferand 550 RPM. As shown in FIG. 27, after being processed for 3 minutes,the resultant morcelized FTSGPs were similarly sized and particledensity as the previous sample after 4 minutes.

In other studies, not included here, similarly morcelized FTSGPs havebeen demonstrated to be injectable through 22 gauge needles. The terminjectable as used herein is meant to include dispensing through asyringe and is not intended to be limited to being injected only intothe body, but also includes dispensing onto the body, such as onto awound.

In a next exemplary morcelization study (test 4 a)—again using the samepatient tissue and processing device and process parameters—multiplelarger portions of tissue, measuring approximately 2 cm×3 cm wereinserted into the processor and morcelized for 4 minutes. FIG. 28A is aclose-up of a portion of the FTSG prior to processing, with a sectionalview revealing the thin layer of epidermal tissue (typically includingpigmented stratum corneum, stratum lucidum, statum granulosum, thicklycell populated stratum spinosum, and stratum basale), over the thickerlayer of generally white dermis (including dermal papilla, stem cellrich hair follicles, sweat glands, capillaries, sensory nerve fibers,sebaceous glands and other dermal components—all contained within anabundance of collagen fibers and connective tissue).

FIG. 28B (still test 4 a) shows the resultant dense mixture ofmorcelized particles (MTPs) suspended in 35 ml of buffer solution,contained within the processor chamber. FIG. 28C shows an enlarged viewof the densely populated tissue particle solution presented in a petridish. FIG. 28D shows an enlarged view of the processed FTSGPs, annotatedto point out that the mixture contains differing amounts of epidermis(pigmented) versus dermis (generally whiter), varying proportionally asanticipated looking at the sectional view of pre-processed tissue. Italso appears that the epidermal tissue (more densely populated withcellular structure) slices more sharply, relative to the more fibrousdermal tissue. The epidermal and dermal tissue particles appear to bedistributed rather consistently throughout the mixture.

FIG. 29A were processed using the same device and same processparameters as FIGS. 28A-D, however, on a different day and withabdominoplasty derived tissue from another patient. Together, thesestudies demonstrate a repeatable process, able to achieve consistentFTSGPs outputs, relative to qualitative appearance and particle size, aswell as, consistently high quantitative cellular viability.

A similarly sized (slightly larger) single portion of FTSG, measuringapproximately 2.5 cm×4.5 cm was morcelized for 4 minutes before takingthe photo for FIG. 29A (a different sample test 1b). FIG. 29B (sampletest 1c) was then morcelized for an additional 3 minutes, for a total of7 minutes. And FIG. 29C (sample 1d) was morcelized an additional 3minutes, for a total of 10 minutes. Only a small shallow puddle ofresultant FTSGPs is shown in each of these images so as to bettervisualize individual particle sizes. The overall volume of processedFTSGPs for this study appeared as densely populated as in the previousstudy for FIGS. 28 A-D.

The tabled data in FIG. 25 demonstrates relative consist and repeatablecellular viability outputs for each of the FTSGPs mixtures documentedfor exemplary test samples included in FIGS. 26A-B, 27 and 29A-E.

FTSGPs shown in FIG. 29C (test 1d) above and a subsequent sample (test2b) were further centrifuged in 1.5 ml aliquots for 4 minutes at 700RPM. The resultant tissue form, shown in FIG. 29D demonstrates theability to achieve a fine paste-like mixture which can be dispensedthrough a syringe as demonstrated in FIG. 29E. Such a FTSGP tissue formmay be easily applied and dispersed, for example, over expansive woundsurfaces.

Articular Cartilage Process Verification Studies

Articular cartilage was harvested from the peripheral edges of a bovineknee condyle using a 2.5 mm ring curette and then morcelized inaccordance with the methods disclosed herein and using the apparatus andsystems disclosed herein.

The harvested cartilage portions, shown in FIG. 30A, (test 4a on Jan.16, 2017) ranged in approximate size from about 1.0-2.2 cm long, 2-2.5mm wide and 0.75-1.2 mm thick. The portions of cartilage were inserted3-4 at a time into 35 ml of buffered saline solution, within anapparatus as described previously in FIGS. 1-4 without baffles, with theslicing blades rotating at about 550 RPM within the morcelizingmechanism. The cartilage tissue grafts were morcelized (MTPs) for atotal duration of 15 minutes at room temperature.

The resultant cartilage morcels are shown in FIG. 30B, demonstrating theability to also finely morcelize articular cartilage within a device andby a process as described herein to similarly process full thicknessskin graft tissue.

Further details of the present invention are shown and describedhereinbelow with respect to FIGS. 31-38. These details include thetechnology advantages and components, the needs and benefits, thetechnology procedure, tissue types and preparation, the process,variable tissue particle sizes, tissue dispensing options, clinicalindication and development status.

It is contemplated that the present invention meets a significant unmetneed. Full thickness skin grafts are the gold standard for chronicwounds and burns, but are rarely used because dermatomas create donorsites that do not heal and the procedure must typically be done in anoperating room.

The present invention can generate a full thickness skin graft rapidlywithout leaving a conventional donor site to heal.

The present invention can also be customized to be applied to fit woundanatomy and can be done as an office procedure. The resulting process ofthe present invention and the grafts produced thereby are fast toprocess, are minimally invasive, antiseptic, provide superior viabilityand are cost effective solutions for wound healing. The system,equipment and process of the present invention can be conducted atbedside, including preparation of the morcelized TPs (MTPs) andformation of a fluid having a pH to help sustain the tissues, anddispensing of the MTPs onto/into the area intended to be treated, whichmay be a wound, a cosmetic or plastic surgery area, an internal organarea and the like.

As shown in FIG. 31, use of the device of the present invention, whichincludes a processing device 950, an applicator 952 and reusableequipment 954, allows for retention of the original tissue structure,high tissue/cell viability (90-95%) and the ability to vary tissueparticle size. In addition, versatile dispensing methods such as spreadpaste, spray and injectables may be used. These are all acceptable forin-office procedures and may be completed within approximately 20minutes or less, desirably about ten minutes or less. Moreover, thepresent invention allows for processing of multiple tissue types such askin, cartilage and organs.

Referring to FIG. 32, the complete total procedure is completed withinthirty minutes. Preparation 955 is improved as the procedure results infast healing, low pain levels, fast harvesting and processing, and asuture closed donor site. The processing 956 to form the MTPs isconducted in a closed antiseptic system taking no more than about tenminutes. The process is automated and can accommodate variable particlesize and results in high cell viability (90%+, such as 99%). Application957 may be done by selectable tips on irregular surfaces and withvariable wound sizes. Also, the application may be injectable. The MTPsof the present invention are desirably prepared in a pH suitable formaintaining viability once they have be morcelized into the intendedsizes. The fluid containing the morcelized highly viable MTPs may bedispensed using a conventional syringe onto or into the area to betreated. The fluid containing the MTPs suspended therein may be appliedto a wound, or other area of the body in need of treatment, such as in abody joint, a plastic surgery application or cosmetic application, orother area of use to enhance the health the of tissue and/or overallappearance and health of the patient.

Turning now to FIG. 33, therein as shown, the preparation process usingboth full thickness skin grafts (FTSGs) and cartilage grafts (CG).

FIG. 34 shows the basic three-step process with respect to morcelizedabdominoplasty tissue in solution dispensed from a 1 mm syringe. Thisincludes introduction 958, morcelization (cutting the donor site tissuein particles) 959, and dispensing 961.

FIG. 35 shows variable tissue particle sizes, which may be formed by theinventive process and using the devices and systems discussed herein, offull thickness skin graft particles (FTSGs) (also referred to as morcelsor MTPs) containing cells and extracellular components.

FIG. 36 shows various dispensing tip options and devices includingspreading 980 using a fan-tip wiper 981; a paste 982 using a cannula983; spray 984 using a nozzle 985 and an injectable 986 using a needle987, all coming from an appropriate applicator device 952.

FIG. 37 shows non-limiting examples of clinical indications includingwound healing 990, skin anesthetic injectable 991 and cartilage repair992 using injection devices with appropriate tip selection.

FIG. 38 shows morcelized tissue particles (MTPs) of bovine kneearticular cartilage, as well as morcelized tissue from abdominoplasticformed using the inventive process. These morcelized cartilage and skinparticulates (particles) may be disbursed to a patient using any of thedispensing devices described herein. These results had been repeatedlyverified to have 87-98% cellular viability using standard tripan and MTTtest protocol.

Further embodiments of the present invention are now described withrespect to FIGS. 39-50 where similar description and reference numeralsare used to describe similar components.

FIG. 39 shows a processing device 1010 having a processing container orchamber 1012 which is similar to that described above. In addition,processing device 1010 also includes a filter chamber 1100, a drainchamber 1200 and a dispenser 1300. The processing chamber 1012 is filledwith a measured volume of sterile solution, preferably sterile saline.The TBS is inserted into the processing chamber 1012 through an upperopen end 1014 and into the solution. The sterile solution and TBS isshown collectively as tissue fluid (suspension) 1018. The processingchamber 1012 may include a cover or lid 1016 for closure of the open end1014. The lid 1016 may be detachable or preferably hinged upon theprocessing chamber 1012 to cover the open end 1014 during processing.

In the present embodiment, the morcelizing mechanism 1040 is driven byan axially rotating drive shaft 1042 that passes through a suitablerotary shaft seal 1047 that may be a silicone part held in place by aretaining clip 1048 that prevents fluid from leaking from the processingchamber 1012.

Processing chamber 1012 further includes baffle panels 1050 within theinterior thereof. With additional reference to FIGS. 49 and 50, thebaffle panels 1050, arrayed about a center axis, are preferably integralwith the interior wall 1013 of the processing chamber 1012 or mayadjacently abut the interior wall. The baffle panels 1050 enhance flowand circulation characteristics of the MTPs (e.g. FTSGs) within thefluid through the morcellation mechanism 1040, by disrupting otherwisecircumferential flow to effectively divert flow into and through themorcelizing mechanism 1040.

Referring again to FIG. 39, fluid communication is provided betweenprocessing chamber 1012 and filter chamber 1100 through a connectingchannel 1416. A flow valve 1417 positioned within connecting channel1416 is shown in the closed position. The valve could be a ball valve,choke valve, stopcock or any other means to controllably open/closefluid flow from the processing chamber 1010 into the filter chamber 1100and further on into the drain chamber 1200.

The drain chamber 1200 mentioned above is shown as an annular shapedcontainer.

FIG. 40 shows the processing device of FIG. 39 with the flow valve 1417in the open position. Fluid communication between the processing chamber1012 and the filter chamber 1100 through connecting channel 1416 enablesthe fully processed MTPs in fluid suspension 1019 to drain into thefilter chamber 1100. Upon completing morcelization, as discussed above,the morcelizing mechanism 1040 continues to rotate to maintain the MTPsin suspension as the MTPs and fluid 1019 drain through the opened valve1417 and into an inner chamber 1410 of the filter chamber 1100 indicatedby arrow A.

The drain chamber 1200 can include a vent (not shown) to enable airwithin the head space to escape ahead of incoming fluid flow when theflow valve 1417 is opened.

Additionally and as more fully shown in FIG. 48, within the filterchamber 1100, a cylindrical filter 1402 separates the inner chamber 1410from an outer chamber 1420. Ribs 1430 protruding from the inner surfacesof the walls of filter chamber create linear drain channels 1435 andalso support the cylindrical filter 1402. The structure of thecylindrical filter may be interrupted locally for flow from theconnecting channel 1416 into the inner chamber 1410 of the filterchamber 1100. Alternatively, the connecting channel 1416 may pass overthe filter 1402 into the inner chamber of the filter chamber. The outersurface of the filter 1402 is supported by the array of vertical ribs1430 that run lengthwise between the cylindrical filter 1402 and insidethe outer filter chamber 1420. Spaces between the ribs 1430 form thedrain channels 1435 that enable the filtrate 1023 to pass through thefilter and drain down the peripheral drain channel 1435 into the drainchamber 1200 below the filter chamber 1100 as indicated by arrows B inFIG. 40. The drain channel 1435 may alternatively drain the filtrate toa dispensing port 1415 to void or collect the filtrate externally fromthe device.

The cylindrical filter 1402, which may or may not form a complete 360degree cylinder, permits fluid and particles smaller that the filterpore size (mesh size) to drain through the filter and down the drainchannels 1435 and out through a lower filter chamber drain port 1440(FIG. 40) communicating with the drain channels 1435 into the drainchamber 1200.

The filter 1402 mentioned above can be formed into a cylinder from aflat filter perforated sheet such as a stainless-steel sheet withphotochemically etched openings or for example lasercut perforationsthrough a plastic sheet. The cylindrical surface area of the filter islarge enough and with sufficient pore density to prevent MTPs from fullyoccluding the filter. MTPs settle and collect on the inner surface ofthe filter and settle on the inner concave bottom 1411 of the filterchamber 1100. The filter 1402, separates the inner filter chamber 1410from the outer filter chamber. The filter 1402 serves as a sieve tocollect MTPs which are larger than the filter perforations and to expelexcess solution and filtrate particles smaller than the perforationsfrom the mixture.

A piston 1306 passes through the lumen of the inner filter chamber 1410and is attached to a plunger 1304 which extends upward and through afilter cap 1308. The cap 1308 and the filter chamber 1410 aremechanically secured via threads, although could be secured by variousother means such as snap features, plastic welds, or adhesive.

The drain chamber 1200 may or may not form a 360-degree annularenclosure. The drain chamber 1200 may be any type of container or meansof collecting the filtrate. An absorbent, such as a compound or anabsorbent material, may be included inside the drain chamber to congeal,coagulate or otherwise increase the viscosity of the filtrate.

Referring more specifically to FIG. 41, the processing device 1010 ofFIGS. 39 and 40 is shown but with the plunger 1304 in the down positionand the filtered MTPs 1021 transferred into the dispenser 1300. Thedownward movement of the plunger 1304 (indicated by arrow C) causes thepiston 1306 to transfer MTPs 1021 that have adhered to the inner wall ofthe filter 1402 and onto the inner concave bottom 1411 of the filterchamber 1100 where other MTPs have also settled. As the piston 1306advances through the filter chamber 1100 and approaches an inner concavebottom 1411. MTPs are transferred from the filter chamber 1410, througha dispensing port 1415 and into the receiving dispenser 1300. As MTPsare driven by the piston 1306 into the receiving dispenser 1300, asyringe plunger 1360 within the dispenser 1300 will be displaced outwardfrom the dispenser. The port 1415 may be a standard type threaded Luerconnector. The dispenser 1300 may be a standard type syringe 1301attached onto the dispenser port 1415 of the filter chamber 1410 or anyother means to collect the MTP matrix. The dispenser containing the MTPsmatrix is removable for dispensing the MTPs, for example onto thetreatment site.

Referring now to FIG. 42, an embodiment similar to FIG. 39 is shownwhere the flow valve is replaced instead by a simple connecting channel1450 between the processing chamber 1012 and the filter chamber 1100.The head pressure of fluid 1018 within the processing chamber 1012 isinitially unable to drive the fluid through the connecting channel 1450,constrained in a state of equilibrium upon the air entrapped belowwithin the interconnected filter chamber 1100 and the drain chamber1200.

Similarly to the embodiment of FIGS. 39-41, the embodiment of FIG. 42,also includes a piston 1306, attached to a plunger 1304 that passesthrough a filter cap 1308. However, here, a plunger seal 1307 within thefilter cap 1308 prevents air within the filter chamber 1100 and drainchamber 1200 from escaping to atmosphere alongside the plunger shaft1304. The seal 1307 may be an elastomeric disk, for example lowdurometer silicone, with a hole through which the plunger passes and iscaptured and held in compression, between the filter cap 1308 andchamber 1100. The cap 1308 is secured onto the filter chamber 1410, forexample via threads, snap features, plastic welds, or adhesive.

A vent valve 1452 is positioned in the upper surface of the drainchamber 1200. The filter chamber 1100 and the drain chamber 1200maintain an airtight enclosure while the vent valve 1452 isspring-loaded closed. Fluid is contained by the processing chamber 1012with a fluid level that is above the connecting channel 1450 thatcommunicates between the processing chamber 1012 and the filter chamber1100. The sealed filter chamber 1100, drain chamber 1200 and connectingchannel 1450 maintain a static volume of air that prevents the fluid1018 within the processing chamber 1012 from flowing into the filterchamber 1100. The connecting channel 1450 restricts fluid communicationbetween the processing and filter chambers such that fluid and aircannot flow through the connecting channel 1450 while the device is atrest and the vent valve 1452 is closed in its static position.

FIG. 43 shows the same device as in FIG. 42 only the vent valve 1452 onthe drain chamber 1200 is now shown open, enabling head pressure of thefluid within the processing chamber 1012 to drive air inside the filterand drain chambers 1100 and 1200 to exit out through the vent valve1452. In this manner the expelled air is displaced by the fluid enteringthe filter chamber, as the MTP are collected within the filter chamber,and the sieve function of the filter enables the fluid and filteredfiltrate 1023 to then flow further to be collected in the drain chamber.

FIG. 44 is similar to the FIG. 42 embodiment showing that the airpressure inside the drain chamber 1200 and the filter chamber 1100similarly prevents the tissue fluid 1018 in the processing chamber 1012from draining into the filter chamber 1100 until a vent valve 1454 isopened. The vent valve 1454 in FIG. 44 is shown in a closed state. Thisvent valve 1454 is configured to be actuated from the bottom of thedevice such that the opening of the valve 1454 can be automatedelectromechanically. A valve seal 1456 is located inside the upperregion of the drain chamber, well above the maximum level that entersthe drain chamber.

The device of FIG. 44 is shown in FIG. 45 with the vent valve 1454 inthe open position. An actuator shaft 1458 is shown extending upward froman operatable processor 1480 (shown in dotted lines), to drive the ventvalve 1454 into the up/open position. The MTPs 1019 have drained intothe filter chamber 1100 and the filtrate 1023 has passed through thefilter 1402 and drained into the drain chamber 1200.

FIG. 46 shows a processing device similar to FIG. 44 in that it isconfigured to be actuated from below from the operatable processor 1480.This figure shows the device configured to provide for automation of alinearly actuated flow valve 1490 between the processing chamber 1012and filter chamber 1100, as well as a linearly actuated plunger 1492within the filter chamber.

In this embodiment, the linear actuated valve 1490 would be closed (FIG.47) in the initial state when fluid and TBSs are contained within theprocessing chamber for processing. As shown in FIG. 46, the valve 1490is then driven to an open state by an actuator shaft 1491 followingmorcelization, to enable fluid to drain into the filter chamber 1100 andpass through into the drain chamber 1200, while the MTPs 1019 drain intothe inner filter chamber 1410 and settle above the piston 1493.

In this embodiment, the linearly actuated piston 1493 is positioned inthe bottom of the filter chamber 1100 during processing, as well as uponopening the flow valve 1490. With use of the automated operableprocessor 1480, upon fully emptying the processing chamber, the motordrive in the operatable processor 1480 will be automatically stopped andthe piston 1493 will then be automatically raised to drive the MTPscaptured within the filter chamber 1100 up into a detachably affixeddispenser 1300.

A linearly positionable piston raising rod 1492 may be adjoined by aseal (not shown) to prevent leakage from the filter chamber 1100. Theflow valve 1490 and/or the valve actuating rod may similarly be adjoinedby a sealing means to prevent leakage from the connecting channel 1450.

FIG. 47 depicts a detail of the processing device in FIG. 46 with thepiston 1493 fully translated to its second state transferring the MTPsfrom the inner filter chamber 1410 into a dispenser 1300. The dispenser1300 can be removed to dispense the MTPs 1021.

Referring again to FIGS. 49 and 50, the baffle panels 1050 mentionedabove are further shown. The morcelizing mechanism 1040 is visible inthe cross section of FIG. 49. The baffle panels 1050 enhance the flowpatterns within the processing chamber and increase the recirculation ofMTPs through the morcelizing mechanism. Preferably there are threebaffle panels, however, there could be more or less. The baffle panels1050 of FIG. 49 are shown preferably integral to the processing chamberwall 1013, facilitating blended surface transitions for effective fluidflow and to avoid such sharp edge features as might otherwise causecirculating MTPs to become hung-up. Alternatively, however, the bafflepanels 1050 could be separate from the wall 1013 and may have spacebetween the processing chamber wall 1013 and the baffle walls 1050 (asshown, for example, in earlier FIG. 20). The baffles of the panels 1050can vary in shape, height, and geometry, and may have perforations oropenings in the baffle panels 1050.

Further embodiments of the blade/cutting assembly 1699 used in themorcelizing mechanism 1040 of the present invention are shown in FIGS.51-57.

FIGS. 51, 51A and 52 show a rigid blade 1700 that is held in compressionby a spring 1710 (Belleville washer) to a cutting disk 1720 and a pin1730 using a retaining clip 1740. The blade 1700 has sharpened cuttingedges 1711 on opposite ends of the blade such that both cutting edgesare in the direction of rotation. The cutting edges 1711 preferably arestraight but could alternatively be curved within the SP. An impeller1717 drives the rotation of the blades. Vertical contact surfaces 1719on the impeller 1717 engage with rear vertical flat engagement surfaces1721 on either side of the blade 1700 to drive the blade rotationallyabout a central axis as the impeller 1717 rotates.

The blade 1700 has upper flat surfaces 1712 that ride in the SP andagainst the disk. The upper flat surfaces 1712 could also be offset atan angle relative to the SP, creating a relief angle, so that only theedge of the blade is in the SP and in contact with the disk 1720.

The cutting disk 1720 has preferably three cutting apertures or breaches1760 which are radially arrayed about a central axis. The blade 1700preferably includes two opposed cutting edges 1711 projecting radiallyoutward about a central axis. The blade 1700 is mounted about the axispin 1730, such that the cutting edges 1711 of the blade 1700 are in theshear plane (SP) side of the cutting disk 1720.

The cutting disk 1720 is a stationary cutting member similar to thatdescribed above. A mating flat or flats 1731 on the axis pin 1730 and onthe central axis hole 1722 through the disk 1720 prevent the disk fromrotating upon the axis pin. An enlarged diameter step 1732 on the axispin 1730 controls axial movement of the disk upon the axis pin.Similarly, the pin has two opposing flat surfaces 1734 that mate intokeyed surfaces in a central hole within the processing chamber. Theretaining clip 1740, for example an ‘E-clip’ or ‘C-clip’, is engagedonto an annular groove about the axis pin and controls the axialmovement of the blade 1700 upon the axis pin 1730 while leaving theblade free to rotate upon the pin 1730.

One or more washers 1733 may be placed between the blade 1700 andretaining clip 1740 to prevent the retaining clip 1740 from beingdislodged as the blade rotates, as well as to prevent the retaining clipfrom wearing against the rotating blade 1700. One or more springs, forexample Bellville washer 1710 or a wave washer, are positioned betweenthe blade 1700 and washers 1733, axially opposed between the retainingclip 1740 and the disk 1720, against the step on the axis pin 1732. Theblades 1700 are secured in compression against the shear plane (SP) sideof the disk 1720.

Another embodiment of the blade/cutting assembly 1699 shown without theimpeller in FIGS. 53-54 is a combination of the morcelizing mechanism 40in FIGS. 5-12 and the morcelizing mechanism 1040 in FIGS. 51-52. Therigid beam 1800 of FIG. 53-54 supports two blades 1810 (or cutting blademembers) on either side of the rigid beam and replaces the blade 1700 inFIG. 51. Each blade 1810 includes a cutting edge 1811. As shown in theblade assembly 1699 in FIGS. 53 and 54, the rigid beam 1800 andsupported blades 1810 are held in compression against a cutting disk1720 by a spring 1710 held together by a retaining clip 1740 and a pin1730 that keys into the disk 1720. The blade retaining features of therigid beam 1800 in FIGS. 53-54 are similar to the blade retainingfeatures on the impeller 108 in FIGS. 5-11. The rear portion 1815 of theblade 1810, opposite of the cutting edge 1811, is secured to the rigidbeam 1800 by screws 1817 in lieu of a slotted feature in the impeller108 shown in FIGS. 5-7 and FIGS. 9-11. The blade could be secured to therigid beam by other means such as a slotted groove, heat stake, a pin orpins. The rigid beam 1800 and blades 1810 are driven rotationallyagainst radially facing flats 1821 by an impeller as described above.The blades 1810 may also be mounted to the rigid beam 1810 such thatthey are parallel and flush against the SP surface of the disk 1720. Theblades 1800 preferably have a straight edge although the edge could becurved, or circular. The blade edges 1811 are held against the SP of thedisk 1720 in compression.

FIG. 55 depicts a blade assembly similar to that shown in FIGS. 53-54,except blades 1811 are thin and can elastically deform or bend incompression against the cutting disk 1720. The blade, being slightlyflexible, may bend into the aperture openings 1760 in the cutting disk1720 such that the cutting edges of the blade ride against the cuttingedges of the cutting disk.

FIG. 56 shows output of a FTSG processed in accordance with the presentinvention for 3 minutes into approximately 1 ml output volume ofmorselized FTSGPs, with each morsel generally 1-3 mm in size. Smallermicro-particle filtrate has been filtered out of the mixture. For scalereference, the morselized FTSGPs of FIG. 56 are shown within a 9 cmpetri dish alongside a metric scale. The morsels are shown diluted insolution to distribute the morsels over the area within the petri dish.

FIG. 57 shows half of the total 1 ml output volume of FTSGPs of FIG. 56,diluted in solution within a 9 cm diameter petri dish. The morsels areagain generally spaced apart over the area of the petri dish, in thisexample illustrating a distribution of morsels at 50% density relativeto the full output volume shown in FIG. 56.

FIG. 58 shows a quarter of the total output volume of FTSGPs of FIG. 56,again diluted in solution within a 9 cm diameter petri dish. In thisexample, the generally spaced apart morsels illustrate a distribution ofmorsels at 25% density relative to the full output volume shown in FIG.56.

Referring now to FIG. 59, an excised elliptically shaped FTSG, is shownhere for scale reference in a 9 cm diameter petri dish (shown in partialview). Placed upon a 1 cm scale grid, the excised FTSG is seen to beapproximately 1 cm×2 cm. FIG. 60 shows an abundance of FTSGPs, also in a9 cm diameter petri dish, processed for 4 minutes from the FTSG of FIG.59 in accordance with the present invention.

FIG. 61 shows a larger excised elliptical FTSG measuring approximately 5cm×2.5 cm, relative to the scale below. FIG. 62 shows an exponentiallylarger output volume of FTSGP, similarly processed for 4 minutes, fromthis larger FTSG. The compared FTSG input and FTSGP output volumes shownin FIGS. 59 and 60, relative to FIGS. 61 and 62, illustrate, by way ofexample, how a desired output volume of processed FTSGPs is easilyachieved by varying the input volume of FTSG.

FIG. 63 shows finely morselated cartilage. Varying the processing time,for example from 4 minutes to 12 minutes, enables precise andreproducible control of output morsel size. In this example, a smallbiopsy sample of cartilage has been processed to output an abundance ofsmall morsels, allowing for broad area coverage. Precisely reproduciblesmall morsels may be delivered by syringe or through an arthroscopicportal.

FIG. 64 shows a planar slice through a single particle of morselizedcartilage using fluorescent confocal microscopy. Staining with calceinAM reveals viable cells as green and dead cells as red. Both green andred are shown together here in this grayscale image. Cartilage morselscontain highly viable tissue components. The live/dead ratio of thisexemplary scan is 88%.

The above-presented description and figures are intended by way ofexample only, and are not intended to limit the present invention in anyway except as set forth in the following claims. It is particularlynoted that persons skilled in the art can readily combine varioustechnical aspects of the elements of the various exemplary embodimentsdescribed above in numerous other ways, all of which are considered tobe within the scope of the invention.

What is claimed is:
 1. A device for processing tissue biopsy samples(TBSs) into morcelized tissue particles (MTPs) comprising: a) aprocessing chamber for accommodating fluid and said TBSs; b) amorcelizing mechanism supported within said processing chamber forrecirculating said fluid and said TBSs in the processing chamber andmorcelizing said TBSs into said MTPs; c) a filter chamber in fluidcommunication with said processing chamber for filtering said fluid andsaid MTPs into a matrix based on a desired size range; d) a drain incommunication with said filter chamber for draining said filtrate andparticles smaller than the desired size range; and e) a dispensing portin communication with said filter chamber to enable collection of thefiltered matrix for subsequent dispensing.
 2. A device of claim 1further including a drain chamber in communication with said drain forcollecting said filtrate and said particles.
 3. A device of claim 1further including a dispensing chamber in communication with saiddispensing port for collection of said filtered matrix.
 4. A device ofclaim 3 wherein said dispensing chamber includes a syringe.
 5. A deviceof claim 3 wherein said processing chamber, said morcelizing mechanism,said filter chamber, said drain chamber and said dispensing port aresterilizable by gamma or electric beam radiation.
 6. A device of claim 1wherein said processing chamber is transparent.
 7. A processing devicefor morcelizing tissue biopsy samples comprising: a processing chamberfor receiving said tissue biopsy samples; an axial pin support in fixedposition within said chamber; a cutting disk non-rotatably mounted onsaid axial pin; a cutting blade rotatably mounted on said axial pin inbiased engagement with said cutting disk; a retaining clip positioned onsaid axial pin for maintaining said cutting blade in said biasedrotational engagement with said cutting disk; and an impeller engagingsaid cutting blade and for rotating said cutting blade about said axialpin and for recirculating said tissue biopsy samples between saidcutting disk and said cutting blade.
 8. A device of claim 7 wherein saidcutting blade includes a pair of opposed cutting edges, said edges beingin biased engagement with said cutting disk.
 9. A device of claim 1wherein said cutting blade includes a pair of cutting blade members eachsupported separately on a rigid beam.
 10. A device of claim 9 whereineach said cutting blade members include a cutting edge held incompression against said cutting disk.
 11. A device of claim 10 whereinsaid blade edges are flexible.
 12. A device of claim 8 wherein saidcutting blade includes engagement surfaces opposite said cutting edgesand wherein said impeller includes contact surfaces for engagement withsaid engagement surfaces.